Journal of virological methods最新文献_第3页

Role of human cytomegalovirus in the pathogenesis of periapical lesions – A systematic review and meta-analysis 人巨细胞病毒在根尖周围病变发病机制中的作用——系统综述和荟萃分析。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-30 DOI: 10.1016/j.jviromet.2026.115359

Puja Chatterjee , Mala Kamboj , Anjali Narwal , Anju Devi , Adarsh Kumar

{"title":"Role of human cytomegalovirus in the pathogenesis of periapical lesions – A systematic review and meta-analysis","authors":"Puja Chatterjee , Mala Kamboj , Anjali Narwal , Anju Devi , Adarsh Kumar","doi":"10.1016/j.jviromet.2026.115359","DOIUrl":"10.1016/j.jviromet.2026.115359","url":null,"abstract":"<div><h3>Background</h3><div>Viruses, especially those of the <em>Herpesviridae</em> family, are increasingly linked to the pathogenesis of periapical lesions. Human cytomegalovirus (CMV), capable of lifelong latency and reactivation under stress or infection, can enhance proinflammatory cytokine release, stimulate osteoclast activity, and drive tissue destruction.</div></div><div><h3>Objective</h3><div>To evaluate the prevalence of CMV DNA and/or RNA in periapical lesions and to evaluate its potential role in their pathogenesis.</div></div><div><h3>Materials and methods</h3><div>A comprehensive search was performed using MEDLINE via PubMed, Google Scholar, Scopus, and ResearchGate up to April 2025, following PRISMA guidelines. Original studies that explored the role of CMV in periapical lesions were included. The risk of bias was evaluated with the modified Newcastle-Ottawa Scale, with results generated in Review Manager 5.4. Meta-analyses were carried out using OpenMeta Analyst.</div></div><div><h3>Results</h3><div>Thirty-one studies were included for qualitative analysis, of which 17 met eligibility for meta-analysis. The pooled prevalence of CMV in periapical lesions was 28 % (95 % CI: 17 %–40 %). Symptomatic lesions showed significantly higher CMV detection compared to asymptomatic cases (OR = 2.67, 95 % CI: 1.44–4.95). Subgroup analysis showed that CMV prevalence varied by detection method, with higher estimates using RT-PCR, indicating assay sensitivity as a major source of heterogeneity.</div></div><div><h3>Conclusion</h3><div>This review highlights that CMV was significantly associated with symptomatic and larger periapical lesions, likely contributing to disease progression via immune modulation, cytokine induction, and viral-bacterial interactions. Further research is warranted to clarify its pathogenic role and explore antiviral strategies in endodontic management.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115359"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimization of a GFP–PAC co-expression cassette–based purification system for efficient construction and rapid screening of recombinant HSV-1 基于GFP-PAC共表达盒的高效构建和快速筛选重组HSV-1纯化系统的优化

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-02-11 DOI: 10.1016/j.jviromet.2026.115364

Mishar Kelishadi , Alijan Tabarraei , Kayhan Azadmanesh

{"title":"Optimization of a GFP–PAC co-expression cassette–based purification system for efficient construction and rapid screening of recombinant HSV-1","authors":"Mishar Kelishadi , Alijan Tabarraei , Kayhan Azadmanesh","doi":"10.1016/j.jviromet.2026.115364","DOIUrl":"10.1016/j.jviromet.2026.115364","url":null,"abstract":"<div><div>Oncolytic viruses (OVs) constitute a promising class of cancer therapeutics engineered to selectively replicate in and lyse malignant cells while sparing normal tissues. Among these, recombinant Herpes Simplex Virus type 1 (HSV-1), generated through targeted deletion of immediate-early (IE) genes, has emerged as a leading candidate, with several vectors currently advancing to phase III clinical trials. Genetic alterations are typically introduced into the HSV-1 genome via homologous recombination using shuttle vectors; however, isolation of recombinant viruses from parental populations by plaque purification remains labor-intensive and technically demanding. This limitation primarily results from the low efficiency of homologous recombination and the frequent persistence of parental virus, which often exhibits a replicative advantage over recombinant variants. In this study, a novel purification protocol incorporating a GFP–PAC co-expression cassette was developed and optimized to enable simultaneous puromycin selection and fluorescence-based identification of recombinant viral constructs. This dual-selection approach significantly accelerates and simplifies the isolation of recombinant HSV-1, enhancing both efficiency and purity. The optimized protocol provides a robust, scalable strategy for generating recombinant HSV-1 with broad applicability in vaccine development, gene therapy, and fundamental virology research.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115364"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146170364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Fully automated sample-to-answer PCR detection of severe fever with thrombocytopenia syndrome virus in cat serum and whole blood 猫血清和全血中发热伴血小板减少综合征病毒的全自动PCR检测。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-24 DOI: 10.1016/j.jviromet.2026.115353

Shun Sasaki , Keita Ishijima , Ken Maeda , Marifu Yamagishi-Kondoh

{"title":"Fully automated sample-to-answer PCR detection of severe fever with thrombocytopenia syndrome virus in cat serum and whole blood","authors":"Shun Sasaki , Keita Ishijima , Ken Maeda , Marifu Yamagishi-Kondoh","doi":"10.1016/j.jviromet.2026.115353","DOIUrl":"10.1016/j.jviromet.2026.115353","url":null,"abstract":"<div><div>Severe fever with thrombocytopenia syndrome virus (SFTSV) is a tick-borne pathogen that causes fatal hemorrhagic disease in humans and animals. In addition to transmission via tick bites, infection can occur through contact with body fluids from infected individuals or animals. This study evaluated the POCKIT™ Central Nucleic Acid Analyzer (POCKIT Central), a fully automated system integrating nucleic acid extraction and insulated isothermal PCR as a rapid diagnostic tool to aid in preventing the spread of SFTSV infection in veterinary medicine. Analytical sensitivity was assessed using four viral strains; all were detectable, although some exhibited reduced sensitivity. In feline whole blood or serum spiked with SFTSV, the system demonstrated sufficient sensitivity for primary diagnosis of cats showing clinical signs of SFTS. Results of analyses of clinical serum samples using POCKIT Central perfectly matched those obtained using conventional reverse transcription–polymerase chain reaction. The POCKIT Central system can detect SFTSV directly in whole blood or serum within 85 min, without requiring complex manual procedures, thus enabling rapid diagnosis and timely therapeutic intervention in veterinary settings, and it also has the potential for application to other animal species.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115353"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146052723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comparative analysis of beet western yellows virus detection methods: Development and validation of a novel real-time RT-PCR assay also targeting beet leaf yellowing virus 甜菜西部黄病毒检测方法的比较分析：一种新的实时RT-PCR检测方法的建立和验证，也针对甜菜叶黄病毒。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-22 DOI: 10.1016/j.jviromet.2026.115352

Hellin Pierre , Everaert Ellen , Demonty Elisabeth , Richet Pauline , Muhovski Yordan , De Jonghe Kris , Hautier Louis , Steyer Stéphan

{"title":"Comparative analysis of beet western yellows virus detection methods: Development and validation of a novel real-time RT-PCR assay also targeting beet leaf yellowing virus","authors":"Hellin Pierre , Everaert Ellen , Demonty Elisabeth , Richet Pauline , Muhovski Yordan , De Jonghe Kris , Hautier Louis , Steyer Stéphan","doi":"10.1016/j.jviromet.2026.115352","DOIUrl":"10.1016/j.jviromet.2026.115352","url":null,"abstract":"<div><div>Beet western yellows virus (BWYV – <em>Polerovirus BWYV</em>) is a polerovirus associated with yellowing and stunting symptoms in sugar beet and several other crops. Its close relationship with other beet-infecting poleroviruses, has long complicated accurate diagnosis due to serological cross-reactivity and limited molecular data. To overcome these issues, we developed a novel real-time RT-PCR assay enabling the specific and sensitive detection of BWYV and beet leaf yellowing virus (BLYV – <em>Polerovirus BLYV</em>). The assay was designed from a curated genomic dataset to ensure inclusivity and tested in parallel with validated reference methods, including a semi-generic ELISA and a generic polerovirus RT-PCR. Comprehensive validation following the EPPO PM 7/98(5) standard confirmed the method’s high analytical sensitivity, specificity, repeatability, and reproducibility. The assay proved robust to methodological variations and compatible with multiplex diagnostic workflows. Compared with existing approaches, it offers faster, more reliable detection and eliminates cross-reactivity with non-target poleroviruses. This method provides a valuable tool for accurate BWYV surveillance and supports improved management of beet virus yellows in agricultural and phytosanitary contexts.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115352"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146044263","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of a strand-specific RT-qPCR assay for detecting and quantifying Borna disease virus RNAs in infected cells 建立一种检测和定量感染细胞中博纳病病毒rna的链特异性RT-qPCR方法。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-25 DOI: 10.1016/j.jviromet.2026.115356

Takehiro Kanda , Shigeki Hattori , Ryo Komorizono , Keizo Tomonaga

{"title":"Development of a strand-specific RT-qPCR assay for detecting and quantifying Borna disease virus RNAs in infected cells","authors":"Takehiro Kanda , Shigeki Hattori , Ryo Komorizono , Keizo Tomonaga","doi":"10.1016/j.jviromet.2026.115356","DOIUrl":"10.1016/j.jviromet.2026.115356","url":null,"abstract":"<div><div>Borna disease virus 1 (BoDV-1) is a nonsegmented negative-strand RNA virus (NSV) that persists in the central nervous system of a broad range of mammalian species. Considering the recent reports linking BoDV-1 to fatal encephalitis in humans, a deeper understanding of its viral life cycle is required. Similar to other NSVs, BoDV-1 produces three types of viral RNAs in infected cells: genome, antigenome, and mRNA. Although a conventional reverse transcription quantitative real-time PCR (RT-qPCR) assay is used to detect viral RNAs, this assay cannot distinguish between RNA species because of primer-independent cDNA synthesis during the reverse transcription (RT) reaction. In this study, we developed a strand-specific RT-qPCR (ssRT-qPCR) assay, in which an RT-primer fused with a non-viral tag sequence at the 5’-end is used in the RT reaction. This assay successfully detected and quantified three types of BoDV-1 RNAs distinctively. Furthermore, using this assay, we measured the intracellular kinetics of the BoDV-1 RNA species, revealing that BoDV-1 may control transcription, but not replication, to establish and maintain persistent infection. Overall, this novel ssRT-qPCR assay is a powerful tool for understanding the life cycle of BoDV-1.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115356"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146064444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A new real-time RT-PCR for the detection of piscine orthoreoviruses PRV-1 and PRV-3 一种检测鱼类正肠病毒PRV-1和PRV-3的实时RT-PCR方法。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-31 DOI: 10.1016/j.jviromet.2026.115361

Laurane Pallandre, Marine Baud, Emma Rolland, Alexiane Talbodec, Laurent Bigarré

引用次数: 0

Genetic influence of IFN-γ gene polymorphisms on hepatitis C progression and recovery IFN-γ基因多态性对丙型肝炎进展和恢复的遗传影响

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-06 DOI: 10.1016/j.jviromet.2026.115337

Khair Rafiq, Sanaullah Khan, Muhammad Bar Khan, Muhammad Yaqoob, Muhammad Adnan

{"title":"Genetic influence of IFN-γ gene polymorphisms on hepatitis C progression and recovery","authors":"Khair Rafiq, Sanaullah Khan, Muhammad Bar Khan, Muhammad Yaqoob, Muhammad Adnan","doi":"10.1016/j.jviromet.2026.115337","DOIUrl":"10.1016/j.jviromet.2026.115337","url":null,"abstract":"<div><h3>Background</h3><div>Hepatitis C virus (HCV) infection is a significant health concern that leads to liver complications. The interferon gamma (INF-γ) and interferon gamma receptor-1 (IFN-γR1) genes play role in viral infections. This study aims to analyse the genetic influences of IFN-γ and IFN-γR1 genes polymorphisms on HCV infection outcomes.</div></div><div><h3>Methodology</h3><div>This cross-sectional study comprised 320 subjects, were classified in to two groups; 160 HCV patients and 160 healthy controls. The HCV patients included 50 subjects with spontaneous viral clearance (SVC), 80 patients with chronic hepatitis C (CHC) and 30 subjects with sustained virological responder (SVR). RNA and DNA were extracted from blood samples. HCV detection and genotyping was performed through PCR. The intron-1 of INF-γ and promoter of IFN-γR1 genes were amplified through PCR and sequenced through Sanger’s method.</div></div><div><h3>Results</h3><div>The frequency of TT, AT and AA genotypes of IFN-γ at + 874 in HCV positive patients was 62(38.75 %), 64(40 %) and 34(21.5 %); in healthy individuals 73(46.0 %), 61(38.12 %) and 26(16.0 %) (p = 0.153). Genotype AT was significantly high in CHC patients 45(57.0 %) while the TT was considerably high in SVC 27(54.0 %) (p < 0.0001). The frequency of TT, TC, and CC genotypes of IFN-γR1 at −56 was 41(25.64 %), 56(35.0 %), and 63(39.36 %) in HCV patients, while 61(38.13 %), 33(20.63 %) and 66(41.25) in healthy controls (p = 0.0001). The TC genotype was detected high in CHC 34(42.60 %) while the CC was observed more in SVC 24(48.0 %) (P < 0.0001).</div></div><div><h3>Conclusion</h3><div>IFN-γ AT and IFN-γR1 TC genotypes are associated with Hepatitis C susceptibility, while IFN-γ TT and IFN-γR1 CC may influence recovery.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115337"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145903961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Reporter assays for HPV16 E1/E1C and E2 mRNA splicing using nanoluciferase and secreted luciferase 利用纳米荧光素酶和分泌荧光素酶检测HPV16 E1/E1C和E2 mRNA剪接的报告基因。

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-29 DOI: 10.1016/j.jviromet.2026.115358

Zixiao Jiang , Yani Zhao , Yunji Zheng , Chengyu Hao , Xiaoxu Cui , Johanna Jönsson , Chengjun Wu , Naoko Kajitani , Stefan Schwartz

{"title":"Reporter assays for HPV16 E1/E1C and E2 mRNA splicing using nanoluciferase and secreted luciferase","authors":"Zixiao Jiang , Yani Zhao , Yunji Zheng , Chengyu Hao , Xiaoxu Cui , Johanna Jönsson , Chengjun Wu , Naoko Kajitani , Stefan Schwartz","doi":"10.1016/j.jviromet.2026.115358","DOIUrl":"10.1016/j.jviromet.2026.115358","url":null,"abstract":"<div><div>We have generated two reporter plasmids that either monitor expression of human papillomavirus type 16 (HPV16) E2 mRNAs using splice site SA2709 or HPV16 E1C mRNAs utilizing splice site SA2582. In plasmid pE2sLuc, the entire E2 open reading frame was replaced by the secreted luciferase (sLuc) reporter gene and in the pE1nLuc plasmid the E1/E1C open reading frame was fused in frame with the nanoluciferase (nLuc) reporter gene. We demonstrate that these reporter plasmids can be used to investigate cis-acting RNA elements and trans-acting factor that regulate splicing to the E2 splice site SA2709 and the alternative E1C- splice site SA2582. Furthermore, we generated a stable C33A cell line carrying HPV16 reporter plasmid pE1nLuc. Using this cell line, we showed that splicing to the HPV16 E2 3’-splice site SA2709 could be perturbed by over expression of SR-proteins or by incubation with a pan-Akt kinase inhibitor. These proteins and substances shifted splicing from E2 splice site SA2709 to the upstream HPV16 3’-splice site SA2582 and enhanced nLuc production, thereby demonstrating the utility of the pE1nLuc cell line as a bioassay for HPV16 E2 mRNA splicing.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115358"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146096838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effects of porcine epidemic diarrhoea virus vaccination timing and route of administration on colostrum antibody levels and prevention of diarrhoea in piglets on a pig farm with porcine epidemic diarrhoea 猪流行性腹泻病毒疫苗接种时间和给药途径对猪流行性腹泻猪场仔猪初乳抗体水平和预防腹泻的影响

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-31 DOI: 10.1016/j.jviromet.2026.115360

Bo Kyu Kang, Dong Suk Min, Deok Il Kim, Seo Hyun Jung, Bo Kyeong Kim, Jin A. Lee, Do Yeon Kim, Jung Young Park, Jin Sik Oh

{"title":"Effects of porcine epidemic diarrhoea virus vaccination timing and route of administration on colostrum antibody levels and prevention of diarrhoea in piglets on a pig farm with porcine epidemic diarrhoea","authors":"Bo Kyu Kang, Dong Suk Min, Deok Il Kim, Seo Hyun Jung, Bo Kyeong Kim, Jin A. Lee, Do Yeon Kim, Jung Young Park, Jin Sik Oh","doi":"10.1016/j.jviromet.2026.115360","DOIUrl":"10.1016/j.jviromet.2026.115360","url":null,"abstract":"<div><div>Piglets rely on passive immunity acquired from maternal-derived antibodies obtained by ingesting colostrum and milk from immunised sows. This study evaluated the differences in colostrum antibody profiles (IgA, IgG, and neutralising antibodies) according to the timing, frequency, and administration route of the porcine epidemic diarrhoea virus (PEDV) vaccination in pigs with confirmed PEDV infection, and to assess the corresponding clinical protective effects of PEDV vaccination in neonatal piglets. Piglets of sows that received two or more vaccinations, including an oral administration of a live attenuated vaccine 1 week before farrowing (3K1L and 5K3K1L groups), showed disappearance of diarrhoea and growth retardation, and exhibited a significantly higher weaning weight. Sows vaccinated with an oral live vaccine approximately 1 week before farrowing had significantly higher IgA levels than those vaccinated 2 weeks before farrowing. Neutralising antibody titres differed among the groups (p = 0.023): the thrice vaccination group (5K3K1L) showed significantly higher titres than the pre-vaccination, once (1K1L and 2K2L), and twice (3K1L) vaccination groups. IgG levels did not differ substantially between the vaccinated and unvaccinated groups. Overall, this study provides scientific evidence linking clinical protection against PEDV to antibody levels, especially IgA, in sow colostrum.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115360"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development and application of a real-time RT-PCR assay for the specific detection of influenza D virus D型流感病毒特异性实时RT-PCR检测方法的建立与应用

IF 1.6 4区医学

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-05 DOI: 10.1016/j.jviromet.2026.115338

Jinfeng Wang , Ruiwen Li , Libing Liu , Jieshi Yu , Ning Li , Xincheng Ji , Yue Yang , Xiaoxia Sun , Wanzhe Yuan , Jianchang Wang

{"title":"Development and application of a real-time RT-PCR assay for the specific detection of influenza D virus","authors":"Jinfeng Wang , Ruiwen Li , Libing Liu , Jieshi Yu , Ning Li , Xincheng Ji , Yue Yang , Xiaoxia Sun , Wanzhe Yuan , Jianchang Wang","doi":"10.1016/j.jviromet.2026.115338","DOIUrl":"10.1016/j.jviromet.2026.115338","url":null,"abstract":"<div><div>Since the influenza D virus (IDV) was first isolated from a swine exhibiting respiratory disease symptoms in the United States in 2011, it has been detected in various animals across more than 20 countries worldwide, including cattle, pigs, goats, sheep, and camels. IDV has emerged as a significant pathogen associated with bovine respiratory disease complex (BRDC). In this study, specific primers and a TaqMan probe were designed based on the conserved region in the PB1 gene to develop a highly specific and efficient real-time RT-PCR assay (RT-qPCR) for IDV. The RT-qPCR assay demonstrated excellent specificity for IDV, without cross-reactions with other common BRDC-associated pathogens. Using <em>in vitro</em> transcribed PB1 RNA as a template, the assay's limit of detection was established at 100 copies/reaction. Furthermore, the developed RT-qPCR assay was evaluated using 417 cattle nasal swab samples collected from 10 different regions in Hebei Province, China, between 2021 and 2022. The results showed that 33 samples (7.91 %) tested positive for IDV. The IDV HEF gene was amplified and sequenced in these 33 positive samples, yielding 10 HEF gene sequences that all belonged to the D/Yama2019 lineage, with nucleotide homology ranging from 98.7 % to 99.1 %. This study successfully developed an RT-qPCR assay for the specific and sensitive detection of IDV, which performed well on the cattle nasal samples. Additionally, it is the first to demonstrate the presence of IDV in cattle herds in Hebei Province, China.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"342 ","pages":"Article 115338"},"PeriodicalIF":1.6,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0